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基于SPR技术的抗CD80生物技术药物结合活性方法的建立

Establishment of anti CD80 drug binding activity method based on SPR technology
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摘要 目的基于表面等离子共振(SPR)技术建立抗CD80生物技术药物结合活性方法,并进行方法学验证和样品测定。方法通过优化试验条件建立合适的结合活性方法,并选择合适的数据分析模型后对建立的方法进行方法学验证。结果通过对各种条件进行筛选及耐用性考察,确定了方法的试验参数:配体固定条件为pH 5.0的醋酸盐缓冲液,固定量为400~5000 RU,结合和解离速率为10~50μL·min^(-1),芯片的再生条件为pH 4.0的枸橼酸缓冲液(10 mmol·L^(-1)枸橼酸钠,100 mmol·L^(-1)NaCl),响应参数为稳态相对响应值(在进样结束后10 s范围内窗口的平均响应值),数据模型为四参数模型。根据以上参数建立了基于SPR技术抗CD80生物技术药物结合活性方法。验证结果如下:线性回归方程的决定系数为0.9994;每个效价水平相对效价测定值的相对偏倚平均值为-0.66%~0.64%,线性回归方程的斜率为0.9867;每个效价水平6次独立测试结果的几何变异系数为0.7%~1.5%,范围为64%~156%。各项验证指标均满足相关验证要求,专属性结果也较好。将建立的结合活性方法应用于抗CD80生物技术药物,测得结合活性相对效价为100%。结论本研究通过优化试验条件建立了基于SPR技术的抗CD80生物技术药物结合活性方法,该方法专属性强、重复性好、准确度高。 Objective Based on surface plasmon resonance(SPR)technology,to established the method of anti-CD80 biotechnological drug binding activity,and to carry out the methodology validation and sample determination of the established method.Methods An appropriate binding activity method was established by optimizing the test parameters,and a suitable data analysis model was selected to validate the established method.Results Through screening various conditions and durability test,the test parameters of the method were determined as follows:the ligand fixation condition was acetate buffer with pH of 5.0,the fixed amount was 400-5000 RU,the binding and dissociation rate was 10-50μL·min^(-1),the chip regeneration condition was pH 4.0 citrate buffer(10 mmol·L^(-1) sodium citrate,100 mmol·L^(-1) NaCl),the response parameter was the relative response value of steady state(the average response value within 10 seconds after the end of the injection),and the data model was a four-parameter model.According to the above parameters,an anti-CD80 biotechnological drug binding activity method based on SPR technology was established.The verification results are as follows:the determination coefficient of the linear regression equation was 0.9994,the average relative bias of the relative potency determination value of each potency level was 0.66%-0.64%,the slope of the linear regression equation was 0.9867,the geometric coefficient of variation of six independent tests for each potency level was 0.7%-1.5%,and the range was 64%-156%.All the verification indicators met the relevant verification requirements,and the specificity results were very well.The established binding activity method was applied to anti-CD80 biotechnological drugs,and the relative potency of binding activity was measured to be 100%.Conclusion In this study,a method of anti CD80 biotechnology drug binding activity based on surface plasmon resonance technology is established by optimizing the test conditions,and this method has strong specificity,good repeatability
作者 董闪闪 史卓维 段徐华 邵泓 陈钢 Shan-Shan DONG;Zhuo-Wei SHI;Xu-Hua DUAN;Hong SHAO;Gang CHEN(Shanghai Institute for Food and Drug Control,NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies,Shanghai 201203,China;Institutes of Biomedical Sciences,Fudan University,Shanghai 200032,China)
出处 《中国药师》 CAS 2023年第10期74-81,共8页 China Pharmacist
基金 上海市“科技创新行动计划”仪器共享配套操作与应用技术研究课题(21142202900) 上海市研发公共服务平台项目(19DZ2294600)。
关键词 表面等离子体共振 抗CD80生物技术药物 结合活性 CM5芯片 Surface plasmon resonance Anti-CD80 biomedicines Binding activity CM5 chip
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